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Knockout Tested Rabbit Recombinant Monoclonal RBCK1 antibody. Carrier free. Suitable for WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples.

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Images

Western blot - Anti-RBCK1 antibody [EPR28157-53] - BSA and Azide free (AB309105), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-RBCK1 antibody [EPR28157-53] - BSA and Azide free (AB309105), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-RBCK1 antibody [EPR28157-53] - BSA and Azide free (AB309105), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-RBCK1 antibody [EPR28157-53] - BSA and Azide free (AB309105), expandable thumbnail
  • Western blot - Anti-RBCK1 antibody [EPR28157-53] - BSA and Azide free (AB309105), expandable thumbnail

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Constituents: 100% PBS

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
WBICC/IFFlow Cyt (Intra)IHC-PIP
Human
Tested
Tested
Tested
Not recommended
Not recommended
Mouse
Tested
Not recommended
Tested
Not recommended
Not recommended
Rat
Tested
Expected
Expected
Not recommended
Not recommended

Tested
Tested

Species

Human, Mouse, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Expected
Expected

Species

Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Mouse

Dilution info

-

Notes

-

Tested
Tested

Species

Human, Mouse

Dilution info

-

Notes

-

Expected
Expected

Species

Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Human, Mouse, Rat

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Human, Mouse, Rat

Dilution info

-

Notes

-

Associated Products

Select an associated product type

1 products for Alternative Version

    Target data

    Function

    E3 ubiquitin-protein ligase, which accepts ubiquitin from specific E2 ubiquitin-conjugating enzymes, such as UBE2L3/UBCM4, and then transfers it to substrates. Functions as an E3 ligase for oxidized IREB2 and both heme and oxygen are necessary for IREB2 ubiquitination. Promotes ubiquitination of TAB2 and IRF3 and their degradation by the proteasome. Component of the LUBAC complex which conjugates linear ('Met-1'-linked) polyubiquitin chains to substrates and plays a key role in NF-kappa-B activation and regulation of inflammation. LUBAC conjugates linear polyubiquitin to IKBKG and RIPK1 and is involved in activation of the canonical NF-kappa-B and the JNK signaling pathways. Linear ubiquitination mediated by the LUBAC complex interferes with TNF-induced cell death and thereby prevents inflammation. LUBAC is recruited to the TNF-R1 signaling complex (TNF-RSC) following polyubiquitination of TNF-RSC components by BIRC2 and/or BIRC3 and to conjugate linear polyubiquitin to IKBKG and possibly other components contributing to the stability of the complex. Together with OTULIN, the LUBAC complex regulates the canonical Wnt signaling during angiogenesis. Binds polyubiquitin of different linkage types.

    Alternative names

    RanBP-type and C3HC4-type zinc finger-containing protein 1, HBV-associated factor 4, Heme-oxidized IRP2 ubiquitin ligase 1, Hepatitis B virus X-associated protein 4, RING finger protein 54, RING-type E3 ubiquitin transferase HOIL-1, Ubiquitin-conjugating enzyme 7-interacting protein 3, HOIL-1, RBCK1, C20orf18, RNF54, UBCE7IP3, XAP3, XAP4

    Recommended products

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    Knockout Tested Rabbit Recombinant Monoclonal RBCK1 antibody. Carrier free. Suitable for WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples.

    Alternative names

    RanBP-type and C3HC4-type zinc finger-containing protein 1, HBV-associated factor 4, Heme-oxidized IRP2 ubiquitin ligase 1, Hepatitis B virus X-associated protein 4, RING finger protein 54, RING-type E3 ubiquitin transferase HOIL-1, Ubiquitin-conjugating enzyme 7-interacting protein 3, HOIL-1, RBCK1, C20orf18, RNF54, UBCE7IP3, XAP3, XAP4

    Key facts

    Isotype

    IgG

    Form

    Liquid

    Clonality

    Monoclonal

    Immunogen
    • The exact immunogen used to generate this antibody is proprietary information.
    Carrier free

    Yes

    Clone number

    EPR28157-53

    Purification technique

    Affinity purification Protein A

    Concentration
    Loading...

    Storage

    Shipped at conditions

    Blue Ice

    Appropriate long-term storage conditions

    +4°C

    Notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free batch production

    For more information, read more on recombinant antibodies.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    Product promise

    We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

    In the unlikely event of one of our products not working as expected, you are covered by our product promise.

    Full details and terms and conditions can be found here:
    Terms & Conditions.

    7 product images

    • Western blot - Anti-RBCK1 antibody [EPR28157-53] - BSA and Azide free (ab309105), expandable thumbnail

      Western blot - Anti-RBCK1 antibody [EPR28157-53] - BSA and Azide free (ab309105)

      This data was developed using the same antibody clone in a different buffer formulation (abAB309104).

      Western blot: Anti-RBCK1 antibody [EPR28157-53] (Anti-RBCK1 antibody [EPR28157-53] ab309104) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (Anti-Calnexin antibody [CANX/1543] ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-RBCK1 antibody [EPR28157-53] ab309104 was shown to bind specifically to RBCK1. A band was observed at 58 kDa in wild-type A549 cell lysates with no signal observed at this size in RBCK1 knockout cell line. To generate this image, wild-type and RBCK1 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

      All lanes: Western blot - Anti-RBCK1 antibody [EPR28157-53] (Anti-RBCK1 antibody [EPR28157-53] ab309104) at 1/1000 dilution

      Lane 1: Wild-type A549 cell lysate at 20 µg

      Lane 2: RBCK1 knockout A549 cell lysate at 20 µg

      Lane 3: HepG2 cell lysate at 20 µg

      Lane 4: MCF7 cell lysate at 20 µg

      Secondary

      All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

      Performed under reducing conditions.

      Observed band size: 58 kDa

    • Immunocytochemistry/ Immunofluorescence - Anti-RBCK1 antibody [EPR28157-53] - BSA and Azide free (ab309105), expandable thumbnail

      Immunocytochemistry/ Immunofluorescence - Anti-RBCK1 antibody [EPR28157-53] - BSA and Azide free (ab309105)

      This data was developed using Anti-RBCK1 antibody [EPR28157-53] ab309104, the same antibody clone in a different buffer formulation.
      Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labelling RBCK1 with Anti-RBCK1 antibody [EPR28157-53] ab309104 at 1/50 (10.14 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing mostly cytoplasmic staining in HepG2 cell line. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

    • Flow Cytometry (Intracellular) - Anti-RBCK1 antibody [EPR28157-53] - BSA and Azide free (ab309105), expandable thumbnail

      Flow Cytometry (Intracellular) - Anti-RBCK1 antibody [EPR28157-53] - BSA and Azide free (ab309105)

      This data was developed using Anti-RBCK1 antibody [EPR28157-53] ab309104, the same antibody clone in a different buffer formulation.
      Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labelling RBCK1 with Anti-RBCK1 antibody [EPR28157-53] ab309104 at 1/500 dilution (0.1 ug)/Red (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.

    • Flow Cytometry (Intracellular) - Anti-RBCK1 antibody [EPR28157-53] - BSA and Azide free (ab309105), expandable thumbnail

      Flow Cytometry (Intracellular) - Anti-RBCK1 antibody [EPR28157-53] - BSA and Azide free (ab309105)

      This data was developed using Anti-RBCK1 antibody [EPR28157-53] ab309104, the same antibody clone in a different buffer formulation.
      Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling RBCK1 with Anti-RBCK1 antibody [EPR28157-53] ab309104 at 1/500 dilution (0.1 ug)/Red (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.

    • Western blot - Anti-RBCK1 antibody [EPR28157-53] - BSA and Azide free (ab309105), expandable thumbnail

      Western blot - Anti-RBCK1 antibody [EPR28157-53] - BSA and Azide free (ab309105)

      This data was developed using Anti-RBCK1 antibody [EPR28157-53] ab309104, the same antibody clone in a different buffer formulation.
      Blocking and diluting buffer and concentration: 5% NFDM/TBST
      The expression molecular weight observed is consistent with what has been described in the literature (PMID: 31209050, PMID: 27006117).
      The higher MW(57kd) band is the ubiquitylated form of RBCK1(PMID: 31209050).
      Bands below 57kd are cleaved RBCK1(PMID: 27006117).
      The blot of lanes1, 2 and 6-8 were developed using a high sensitivity ECL substrate.
      Exposure time: Lanes1, 2 and 6-8: 180 seconds; Lane3-5: 48 seconds

      All lanes: Western blot - Anti-RBCK1 antibody [EPR28157-53] (Anti-RBCK1 antibody [EPR28157-53] ab309104) at 1/1000 dilution

      Lane 1: Human hypothalamus tissue lysate at 20 µg

      Lane 2: Human spinal cord tissue lysate at 20 µg

      Lane 3: Mouse brain tissue lysate at 20 µg

      Lane 4: Mouse spleen tissue lysate at 20 µg

      Lane 5: Mouse kidney tissue lysate at 20 µg

      Lane 6: Rat brain tissue lysate at 20 µg

      Lane 7: Rat spleen tissue lysate at 20 µg

      Lane 8: Rat kidney tissue lysate at 20 µg

      Secondary

      All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

      Observed band size: 57 kDa

    • Western blot - Anti-RBCK1 antibody [EPR28157-53] - BSA and Azide free (ab309105), expandable thumbnail

      Western blot - Anti-RBCK1 antibody [EPR28157-53] - BSA and Azide free (ab309105)

      This data was developed using Anti-RBCK1 antibody [EPR28157-53] ab309104, the same antibody clone in a different buffer formulation.
      Blocking and diluting buffer and concentration: 5% NFDM/TBST.
      The expression molecular weight observed is consistent with what has been described in the literature (PMID: 31209050, PMID: 27006117).
      The higher MW(57kd) band is the ubiquitylated form of RBCK1(PMID: 31209050).
      Bands below 57kd are cleaved RBCK1(PMID: 27006117).
      In Western blot, anti- H3 antibody (Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade ab176842) loading control staining at 1/100000 dilution.

      All lanes: Western blot - Anti-RBCK1 antibody [EPR28157-53] (Anti-RBCK1 antibody [EPR28157-53] ab309104) at 1/1000 dilution

      Lane 1: HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg

      Lane 2: HepG2 transfected with siRNA specifically targeti RBCK1 whole cell lysate at 20 µg

      Secondary

      All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

      Observed band size: 57 kDa

      Exposure time: 59s

    • Western blot - Anti-RBCK1 antibody [EPR28157-53] - BSA and Azide free (ab309105), expandable thumbnail

      Western blot - Anti-RBCK1 antibody [EPR28157-53] - BSA and Azide free (ab309105)

      This data was developed using Anti-RBCK1 antibody [EPR28157-53] ab309104, the same antibody clone in a different buffer formulation.
      Blocking and diluting buffer and concentration: 5% NFDM/TBST.
      The expression molecular weight observed is consistent with what has been described in the literature (PMID: 31209050, PMID: 27006117).
      The higher MW(57kd) band is the ubiquitylated form of RBCK1(PMID: 31209050).
      Bands below 57kd are cleaved RBCK1(PMID: 27006117).

      All lanes: Western blot - Anti-RBCK1 antibody [EPR28157-53] (Anti-RBCK1 antibody [EPR28157-53] ab309104) at 1/1000 dilution

      Lane 1: A549 (human lu carcinoma epithelial cell)whole cell lysate at 20 µg

      Lane 2: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

      Lane 3: C2C12 (mouse myoblast) whole cell lysate at 20 µg

      Secondary

      All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

      Observed band size: 57 kDa

      Exposure time: 180s

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    Product protocols

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